This invention relates to novel guandine substituted compounds, pharmaceutical compositions, processes for their preparation, and use thereof in treating IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 and NAP-2 mediated diseases.
Many different names have been applied to Interleukin-8 (IL-8), such as neutrophil attractant/activation protein-1 (NAP-1), monocyte derived neutrophil chemotactic factor (MDNCF), neutrophil activating factor (NAF), and T-cell lymphocyte chemotactic factor. Interleukin-8 is a chemoattractant for neutrophils, basophils, and a subset of T-cells. It is produced by a majority of nucleated cells including macrophages, fibroblasts, endothelial and epithelial cells exposed to TNF, IL-1xcex1, IL-1xcex2 or LPS, and by neutrophils themselves when exposed to LPS or chemotactic factors such as FMLP. M. Baggiolini et al, J. Clin. Invest. 84, 1045 (1989); J. Schroder et al, J. Immunol. 139, 3474 (1987) and J. Immunol. 144, 2223 (1990); Strieter, et al, Science 243, 1467 (1989) and J. Biol. Chem. 264, 10621 (1989); Cassatella et al, J. Immunol. 148, 3216 (1992).
GROxcex1, GROxcex2, GROxcex3 and NAP-2 also belong to the chemokine xcex1 family. Like IL-8 these chemokines have also been referred to by different names. For instance GROxcex1, xcex2, xcex3 have been referred to as MGSAxcex1, xcex2 and xcex3 respectively (Melanoma Growth Stimulating Activity), see Richmond et al, J. Cell Physiology 129, 375 (1986) and Chang et al, J. Immunol 148, 451 (1992). All of the chemokines of the xcex1-family which possess the ELR motif directly preceding the CXC motif bind to the IL-8 B receptor.
IL-8, GROxcex1, GROxcex2, GROxcex3 and NAP-2 stimulate a number of functions in vitro. They have all been shown to have chemoattractant properties for neutrophils, while IL-8 and GROxcex1 have demonstrated T-lymphocytes, and basophiles chemotactic activity. In addition IL-8 can induce histamine release from basophils from both normal and atopic individuals GRO-xcex1 and IL-8 can in addition, induce lysozomal enzyme release and respiratory burst from neutrophils. IL-8 has also been shown to increase the surface expression of Mac-1 (CD11b/CD18) on neutrophils without de novo protein synthesis. This may contribute to increased adhesion of the neutrophils to vascular endothelial cells. Many known diseases are characterized by massive neutrophil infiltration. As IL-8, Groxcex1, GROxcex2, GROxcex3 and NAP-2 promote the accumulation and activation of neutrophils, these chemokines have been implicated in a wide range of acute and chronic inflammatory disorders including psoriasis and rheumatoid arthritis, Baggiolini et al, FEBS Lett. 307, 97 (1992); Miller et al, Crit. Rev. Immunol. 12, 17 (1992); Oppenheim et al, Annu. Rev. Immunol. 9, 617 (1991); Seitz et al., J. Clin. Invest. 87, 463 (1991); Miller et al., Am. Rev. Respir. Dis. 146, 427 (1992); Donnely et al., Lancet 341, 643 (1993). In addition the ELR chemokines (those containing the amino acids ELR motif just prior to the CXC motif) have also been implicated in angiostasis. Strieter et al, Science 258, 1798 (1992).
In vitro, IL-8, GROxcex1, GROxcex2, GROxcex3 and NAP-2 induce neutrophil shape change, chemotaxis, granule release, and respiratory burst, by binding to and activating receptors of the seven-transmembrane, G-protein-linked family, in particular by binding to IL-8 receptors, most notably the B-receptor. Thomas et al., J. Biol. Chem. 266, 14839 (1991); and Holmes et al., Science 253, 1278 (1991). The development of non-peptide small molecule antagonists for members of this receptor family has precedent. For a review see R. Freidinger in: Progress in Drug Research, Vol. 40, pp. 33-98, Birkhauser Verlag, Basel 1993. Hence, the IL-8 receptor represents a promising target for the development of novel anti-inflammatory agents.
Two high affinity human IL-8 receptors (77% homology) have been characterized: IL-8Rxcex1, which binds only IL-8 with high affinity, and IL-8Rxcex2, which has high affinity for IL-8 as well as for GRO-xcex1, GROxcex2, GROxcex3 and NAP-2. See Holmes et al., supra; Murphy et al., Science 253, 1280 (1991); Lee et al., J. Biol. Chem. 267, 16283 (1992); LaRosa et al., J. Biol. Chem. 267, 25402 (1992); and Gayle et al., J. Biol. Chem. 268, 7283 (1993).
There remains a need for treatment, in this field, for compounds which are capable of binding to the IL-8 xcex1 or xcex2 receptor. Therefore, conditions associated with an increase in IL-8 production (which is responsible for chemotaxis of neutrophil and T-cells subsets into the inflammatory site) would benefit by compounds which are inhibitors of IL-8 receptor binding.
This invention provides for a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8 xcex1 or xcex2 receptor and which method comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In particular the chemokine is IL-8.
This invention also relates to a method of inhibiting the binding of IL-8 to its receptors in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula (I).
Compounds of Formula (I) useful in the present invention are represented by the structure: 
wherein
Z is cyano, OR11, C(O)NR15R16, R18, C(O)OR11, C(O)R11, or S(O)2R17;
R is any functional moiety having an ionizable hydrogen and a pKa of 10 or less;
R1 is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted C1-10 alkyl; C1-10 alkyl; C2-10 alkenyl; C1-10 alkoxy; halosubstituted C1-10 alkoxy; azide; (CR8R8)qS(O)tR4; hydroxy; hydroxy C1-4alkyl; aryl; aryl C1-4 alkyl; aryloxy; aryl C1-4 alkyloxy; heteroaryl; heteroarylalkyl; heterocyclic, heterocyclic C1-4alkyl; heteroaryl C1-4 alkyloxy; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyclic C2-10 alkenyl; (CR8R8)qNR4R5; C2-10 alkenyl C(O)NR4R5; (CR8R8)qC(O)NR4R5; (CR8R8)qC(O)NR4R10; S(O)3H; S(O)3R8; (CR8R8)qC(O)R11; C2-10 alkenyl C(O)R11; C2-10 alkenyl C(O)OR11; (CR8R8)qC(O)OR12; (CR8R8)qOC(O) R11; (CR8R8)qNR4C(O)R11; (CR8R8)qNHS(O)2R19; (CR8R8)qS(O)2NR4R5; or two R1 moieties together may form Oxe2x80x94(CH2)sOxe2x80x94 or a 5 to 6 membered saturated or unsaturated ring, and wherein the aryl, heteroaryl and heterocyclic containing moieties may be optionally substituted;
q is 0, or an integer having a value of 1 to 10;
t is 0, or an integer having a value of 1 or 2;
m is an integer having a value of 1 to 3;
s is an integer having a value of 1 to 3;
R4 and R5 are independently hydrogen, optionally substituted C1-4 alkyl, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C1-4alkyl, heterocyclic, or heterocyclic C1-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from oxygen, nitrogen or sulfur;
HET is an optionally substituted heteroaryl moiety;
R6 and R7 are independently hydrogen or a C1-4 alkyl group; or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom selected from oxygen, nitrogen or sulfur;
R8 is independently selected from hydrogen or C1-4 alkyl;
R10 is C1-10 alkyl C(O)2R8 
R11 is hydrogen, C1-4 alkyl, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC1-4alkyl;
R12 is hydrogen, C1-10 alkyl, optionally substituted aryl or optionally substituted arylalkyl;
R13 and R14 are independently hydrogen or C1-4 alkyl;
v is 0, or an integer having a value of 1 to 4;
R15 and R16 are independently hydrogen, optionally substituted C1-4 alkyl, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, optionally substituted heterocyclicC1-4alkyl, or R15 and R16 may together with the nitrogen to which they are attached form a 5 to 7 member ring optionally containing an additional heteroatom selected from oxygen, nitrogen, or sulfur;
R17 is C1-4 alkyl, NR15R16, OR11, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC1-4alkyl;
R18 is optionally substituted C1-4 alkyl, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC1-4alkyl;
R19 is C1-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC1-4alkyl, heterocyclic, or heterocyclicC1-4alkyl, wherein the all of these moieties may be optionally substituted;
Rd is NR6R7, alkyl, arylC1-4 alkyl, arylC2-4 alkenyl, heteroaryl, hetroaryl-C1-4alkyl, heteroarylC2-4 alkenyl, heterocyclic, heterocyclicC1-4 alkyl, wherein the alkyl, aryl, arylalkyl, heteroaryl, heteroaryl alkyl, heterocyclic, and heterocyclic alkyl rings may be optionally substituted; 
the Exe2x80x2 containing ring is optionally selected from 
The compounds of Formula (I) may also be used in association with the veterinary treatment of mammals, other than humans, in need of inhibition of IL-8 or other chemokines which bind to the IL-8 xcex1 and xcex2 receptors. Chemokine mediated diseases for treatment, therapeutically or prophylactically, in animals include disease states such as those noted herein in the Methods of Treatment section.
In compounds of Formula (I), W is suitably 
The Exe2x80x2 containing ring, denoted by its point of attachment through the asterix (*) may optionally be present. If if it is not present the ring is a phenyl moiety which is substituted by the R1 terms as shown. The Exe2x80x2 ring may be substituted by the R1 moiety in any ring, saturated or unsaturated, and is shown for purposes herein substituted only in the unsaturated ring(s).
R is suitably any functional moiety which provides an ionizable hydrogen having a pKa of 10 or less, preferably from about 3 to 9, more preferably from about 3 to 7. Such functional groups include, but are not limited to, hydroxy, carboxylic acid, thiol, SR2, OR2, NHxe2x80x94C(O)Ra, C(O)NR6xe2x80x2R7xe2x80x2, a substituted sulfonamides of the formula: NHS(O)2Rb, S(O)2NHRc, NHC(X2)NHRb, or a tetrazolyl; wherein X2 is oxygen or sulfur, preferably oxygen. Preferably, the functional group is other than a sulfonic acid, either directly or as a substituent group on the aryl, heteroaryl, or heterocyclic moiety ring, such as in SR2 or OR2. More preferably R is OH, SH, or NHS(O)2Rb.
Suitably, R2 is a substituted aryl, heteroaryl, or heterocyclic moiety which ring has the functional moiety providing the ionizable hydrogen having a pKa of 10 or less.
Suitably, R6xe2x80x2 and R7xe2x80x2 are hydrogen, C1-4 alkyl, aryl, arylC1-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC1-4alkyl, heteroarylC2-4 alkenyl, heterocyclic, heterocyclic C1-4alkyl, heterocyclic C2-4alkenyl moiety, all of which may be optionally substituted one to three times independently by halogen; nitro; halosubstituted C1-4 alkyl, such as CF3; C1-4 alkyl, such as methyl; C1-4 alkoxy, such as methoxy; NR9C(O)Ra; C(O)NR6R7; S(O)3H; or C(O)OC1-4 alkyl, provided that only one of R6xe2x80x2 and R7xe2x80x2 are hydrogen, but not both.
Suitably, R6 and R7 are independently hydrogen or a C1-4 alkyl group, or R6 and R7 together with the nitrogen to which they are attached form a 5 to 7 member ring which ring may optionally contain an additional heteroatom which heteroatom is selected from oxygen, nitrogen or sulfur. This heteroring may be optionally substituted as defined herein.
Suitably Ra is an aryl, arylC1-4alkyl, heteroaryl, heteroarylC1-4alkyl, heterocyclic, or a heterocyclic C1-4alkyl moiety, all of which may be optionally substituted, as defined herein below.
Suitably, Rb is a NR6R7, alkyl, aryl, arylC1-4alkyl, arylC2-4alkenyl, heteroaryl, heteroarylC1-4alkyl, heteroarylC2-4 alkenyl, heterocyclic, heterocyclic C1-4alkyl, heterocyclic C2-4alkenyl moiety, or camphor, all of which may be optionally substituted one to three times independently by halogen; nitro; halosubstituted C1-4 alkyl, such as CF3; C1-4 alkyl, such as methyl; C1-4 alkoxy, such as methoxy; NR9C(O)Ra; C(O)NR6R7; S(O)3H; or C(O)OC1-4 alkyl. Rb is preferably an optionally substituted phenyl, benzyl, or styryl. When Rb is a heteroaryl preferably it is an optionally substituted thiazole, optionally substituted thienyl, or optionally substituted quinolinyl ring.
Suitably R9 is hydrogen or a C1-4 alkyl, preferably hydrogen. Preferably, when the substituent group is NR9C(O)Ra, then Ra is preferably an alkyl group, such as methyl.
Suitably Rc is hydrogen, alkyl, aryl, arylC1-4alkyl, arylC1-4alkenyl, heteroaryl, heteroarylC1-4alkyl, heteroarylC1-4alkenyl, heterocyclic, heterocyclic C1-4alkyl, or heterocyclic C1-4alkenyl moiety, all of which may be optionally substituted one to three times independently by halogen, nitro, halosubstituted C1-4 alkyl, C1-4 alkyl, C1-4 alkoxy, NR9C(O)Ra, C(O)NR6R7, S(O)3H, or C(O)OC1-4 alkyl. Preferably, Rc is an optionally substituted phenyl.
When R is an OR2 or SR2 moiety it is recognized by one of skill in the art that the aryl ring must, therefore, contain the required ionizable hydrogen. The aryl ring may also be additionally substituted, independently, by one to three groups, which groups may also contain an additional ionizable group, and which include but are not limited to, halogen, nitro, halosubstituted C1-4 alkyl, C1-4 alkyl, C1-4 alkoxy, hydroxy, SH, C(O)NR6R7, NHxe2x80x94C(O)Ra, NHS(O)2Rb, S(O)2NR6R7, C(O)OR8, or a tetrazolyl ring.
In compounds of Formula (I), suitably R1 is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted C1-10 alkyl, such as CF3; C1-10 alkyl, such as methyl, ethyl, isopropyl, or n-propyl; C2-10 alkenyl; C1-10 alkoxy, such as methoxy, or ethoxy; halosubstituted C1-10 alkoxy, such as trifluoromethoxy; azide; (CR8R8)q S(O)tR4, wherein t is 0, 1 or 2; hydroxy; hydroxy C1-4alkyl, such as methanol or ethanol; aryl, such as phenyl or naphthyl; aryl C1-4 alkyl, such as benzyl; aryloxy, such as phenoxy; aryl C1-4 alkyloxy, such as benzyloxy; heteroaryl; heteroarylalkyl; heteroaryl C1-4 alkyloxy; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyclic C2-10 alkenyl; (CR8R8)qNR4R5; C2-10 alkenyl C(O)NR4R5; (CR8R8)q C(O)NR4R5; (CR8R8)q C(O)NR4R10; S(O)3H; S(O)3R8; (CR8R8)q C(O)R11; C2-10 alkenyl C(O)R11; C2-10 alkenyl C(O)OR11; C(O)R11; (CR8R8)q C(O)OR12; (CR8R8)qOC(O)R11; (CR8R8)q NR4C(O)R11; (CR8R8)q NHS(O)2R19; or (CR8R8)qS(O)2NR4R5; or two R1 moieties together may form Oxe2x80x94(CH2)sOxe2x80x94 or a 5 to 6 membered saturated or unsaturated ring: and s is an integer having a value of 1 to 3. The aryl , heteroaryl, and heterocyclic containing moieites may be optionally substituted as defined herein below.
Suitably, q is 0, or an integer having a value of 1 to 10.
When R1 forms a dioxybridge, s is preferably 1. When R1 forms an additional saturated or unsaturated ring, it is preferably 6 membered ring resulting in a naphthylene ring system. These rings may be substituted independently, 1 to 3 times by the other R1 moieties as defined above.
Suitably, R4 and R5 are independently hydrogen, optionally substituted C1-4 alkyl, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl C1-4alkyl, heterocyclic, heterocyclicC1-4 alkyl, or R4 and R5 together with the nitrogen to which they are attached form a 5 to 7 member ring which may optionally comprise an additional heteroatom selected from O/N/S.
R8 is suitably independently selected from hydrogen or C1-4 alkyl.
R10 is suitably C1-10 alkyl C(O)2R8, such as CH2C(O)2H or CH2C(O)2CH3.
R11 is suitably hydrogen, C1-4 alkyl, aryl, aryl C1-4 alkyl, heteroaryl, heteroaryl C1-4alkyl, heterocyclic, or heterocyclic C1-4alkyl.
R12 is suitably hydrogen, C1-10 alkyl, optionally substituted aryl or optionally substituted arylalkyl.
R19 is C1-4alkyl, aryl, arylalkyl, heteroaryl, heteroarylC1-4alkyl, heterocyclic, or heterocyclicC1-4alkyl, wherein the all of these moieties may be optionally substituted;
Preferably R1 is halogen, cyano, nitro, CF3, C(O)NR4R5, alkenyl C(O)NR4R5, C(O) R4R10, alkenyl C(O)OR12, heteroaryl, heteroarylalkyl, heteroaryl alkenyl, or S(O)NR4R5, and preferably R4 and R5 are both hydrogen or one is phenyl. A preferred ring substitution for R1 is in the 4-position of the phenyl ring.
When R is OH, SH or NS2Rb than R1 is preferably mono-substituted in the 3-position, the 4-position or is di substituted in the 3,4-position. The substituent group is suitably an electron withdrawing moiety. Preferably when R is OH, SH or NSO2Rb, than R1 is nitro, halogen, cyano, trifluoromethyl group, C(O)NR4R5.
When R is carboxylic acid, than R1 is preferably hydrogen, or R1 is preferably substituted in the 4-position, more preferably substituted by trifluoromethyl or chloro.
In compounds of Formula (I), suitably R13 and R14 are independently hydrogen, optionally substituted C1-4 alkyl which may be straight or branched as defined herein, or one of R13 and R14 are an optionally substituted aryl; v is 0, or an integer having a value of 1 to 4.
When R13 or R14 are an optionally substituted alkyl, the alkyl moiety may be substituted one to three times independently by halogen; halosubstituted C1-4 alkyl such as trifluromethyl; hydroxy; hydroxy C1-4alkyl, C1-4 alkoxy; such as methoxy, or ethoxy, halosubstituted C1-10 alkoxy, S(O)tR4; aryl; NR4R5; NHC(O)R4; C(O)NR4R5; or C(O)OR8.
In compounds of Formula (I), Z is suitably cyano, OR11, C(O)NR15R16, R18, C(O)R11, C(O)OR11, or S(O)2R17.
Suitably R15 and R16 are independently hydrogen, optionally substituted C1-4 alkyl, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, optionally substituted heterocyclicC1-4alkyl, or R15 and R16 may together with the nitrogen to which they are attached form a 5 to 7 member ring optionally containing an additional heteroatom selected from oxygen, nitrogen, or sulfur.
Suitably, R17 is C1-4 alkyl, NR15R16, OR11, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC1-4alkyl.
In compounds of Formula (I), R18 is suitably hydrogen, halogen, cyano, optionally substituted C1-4 alkyl, halo substituted C1-4 alkyl, C(O)NR15R16, optionally substituted aryl, optionally substituted aryl C1-4alkyl, optionally substituted heteroaryl, optionally substituted heteroarylC1-4alkyl, optionally substituted heterocyclic, or optionally substituted heterocyclicC1-4alkyl.
In compounds of Formula (I), HET is suitably a heteroaryl ring or multi-ring system. If the HET moiety is a multi ring system, the ring containing the heteroatom does not need to be directly attached to the urea moiety. All the rings in this ring system may be optionally substituted as defined herein. Preferably the HET moiety is a pyridyl, which may be 2-, 3- or 4-pyridyl. If the ring is a multi system ring it is preferably benzimidazole, dibenzothiophene, or an indole ring. Other heterocyclic rings of interest include, but are not limited to thiophene, furan, pyrimidine, pyrrole, pyrazole, quinoline, isoquinoline, quinazolinyl, pyridine, oxazole, thiazole, thiadiazole, triazole, imidazole, or benzimidazole.
In compounds of Formula (I), the HET ring may be optionally substituted independently one to three times by Y, i.e. (Y(n)), wherein n is an integer having a value of 1 to 3. Suitably, Y is independently selected from hydrogen; halogen; nitro; cyano; halosubstituted C1-10 alkyl; C1-10 alkyl; C2-10 alkenyl; C1-10 alkoxy; halosubstituted C1-10 alkoxy; azide; (CR8R8)q S(O)tR4; hydroxy; hydroxyC1-4alkyl; aryl; aryl C1-4 alkyl; aryloxy; arylC1-4 alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl C1-4 alkyloxy; heterocyclic, heterocyclic C1-4alkyl; aryl C2-10 alkenyl; heteroaryl C2-10 alkenyl; heterocyclic C2-10 alkenyl; (CR8R8)q NR4R5; C2-10 alkenyl C(O)NR4R5; (CR8R8)q C(O)NR4R5; (CR8R8)q C(O)NR4R10; S(O)3H; S(O)3R8; (CR8R8)q C(O)R11; C2-10 alkenyl C(O)R11; C2-10 alkenyl C(O)OR11; (CR8R8)q C(O)OR12; (CR8R8)q OC(O)R11; (CR8R8)q NR4C(O)R11; (CR8R8)q NHS(O)2Rd; (CR8R8)q S(O)2NR4R5; or two Y moieties together may form Oxe2x80x94(CH2)sOxe2x80x94 or a 5 to 6 membered saturated or unsaturated ring; wherein the aryl, heteroaryl, heterocyclic containing moieties may be optionally substituted as defined herein.
When Y forms a dioxybridge, s is preferably 1. When Y forms an additional saturated or unsaturated ring, it is preferably 6 membered ring resulting in a naphthylene ring system. These rings may also be substituted 1 to 3 times by other Y moieties as defined above.
Suitably, Rd is a NR6R7, alkyl, aryl C1-4 alklyl, arylC2-4 alkenyl, heteroaryl, hetroaryl-C1-4alkyl, heteroarylC2-4 alkenyl, heterocyclic, heterocyclicC1-4 alkyl, or heterocyclic C2-4 alkenyl moiety, wherein the aryl, heteroaryl, and heterocyclic containing moieties may all be optionally substituted as defined herein.
As used herein, xe2x80x9coptionally substitutedxe2x80x9d unless specifically defined shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; hydroxy substituted C1-10alkyl; C1-10 alkoxy, such as methoxy or ethoxy; S(O)mxe2x80x2 C1-10 alkyl, wherein mxe2x80x2 is 0, 1 or 2, such as methyl thio, methyl sulfinyl or methyl sulfonyl; amino, mono and di-substituted amino, such as in the NR4R5 group; NHC(O)R4; C(O)NR4R5; C(O)OH; S(O)2NR4R5; NHS(O)2R21, C1-10 alkyl, such as methyl, ethyl, propyl, isopropyl, or t-butyl; halosubstituted C1-10 alkyl, such CF3; an optionally substituted aryl, such as phenyl, or an optionally substituted arylalkyl, such as benzyl or phenethyl, optionally substituted heterocylic, optionally substituted heterocylicalkyl, optionally substituted heteroaryl, optionally substituted heteroaryl alkyl, wherein these aryl, hetroaryl, or heterocyclic moieties may be substituted one to two times by halogen; hydroxy; hydroxy substituted alkyl; C1-10 alkoxy; S(O)mxe2x80x2C1-10 alkyl; amino, mono and di-substituted amino, such as in the NR4R5 group; C1-10 alkyl, or halosubstituted C1-10 alkyl, such as CF3.
R21 is suitably C1-4 alkyl, aryl, aryl C1-4alkyl, heteroaryl, heteroarylC1-4alkyl, heterocyclic, or heterocyclicC1-4alkyl.
Suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of inorganic and organic acids, such as hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methane sulphonic acid, ethane sulphonic acid, acetic acid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid, phenylacetic acid and mandelic acid. In addition, pharmaceutically acceptable salts of compounds of Formula (I) may also be formed with a pharmaceutically acceptable cation, for instance, if a substituent group comprises a carboxy moiety. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations.
The following terms, as used herein, refer to:
xe2x80x9chaloxe2x80x9dxe2x80x94all halogens, that is chloro, fluoro, bromo and iodo.
xe2x80x9cC1-10alkylxe2x80x9d or xe2x80x9calkylxe2x80x9dxe2x80x94both straight and branched chain radicals of 1 to 10 carbon atoms, unless the chain length is otherwise limited, including, but not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl and the like.
The term xe2x80x9ccycloalkylxe2x80x9d is used herein to mean cyclic radicals, preferably of 3 to 8 carbons, including but not limited to cyclopropyl, cyclopentyl, cyclohexyl, and the like.
The term xe2x80x9calkenylxe2x80x9d is used herein at all occurrences to mean straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl and the like.
xe2x80x9carylxe2x80x9dxe2x80x94phenyl and naphthyl;
xe2x80x9cheteroarylxe2x80x9d (on its own or in any combination, such as xe2x80x9cheteroaryloxyxe2x80x9d, or xe2x80x9cheteroaryl alkylxe2x80x9d)xe2x80x94a 5-10 membered aromatic ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O or S, such as, but not limited, to pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, oxazole, thiazole, thiadiazole, triazole, imidazole, or benzimidazole.
xe2x80x9cheterocyclicxe2x80x9d (on its own or in any combination, such as xe2x80x9cheterocyclicalkylxe2x80x9d)xe2x80x94a saturated or partially unsaturated 4-10 membered ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, O, or S; such as, but not limited to, pyrrolidine, piperidine, piperazine, morpholine, tetrahydropyran, or imidazolidine.
The term xe2x80x9carylalkylxe2x80x9d or xe2x80x9cheteroarylalkylxe2x80x9d or xe2x80x9cheterocyclicalkylxe2x80x9d is used herein to mean C1-10 alkyl, as defined above, attached to an aryl, heteroaryl or heterocyclic moiety, as also defined herein, unless otherwise indicated.
xe2x80x9csulfinylxe2x80x9dxe2x80x94the oxide S (O) of the corresponding sulfide, the term xe2x80x9cthioxe2x80x9d refers to the sulfide, and the term xe2x80x9csulfonylxe2x80x9d refers to the fully oxidized S(O)2 moiety.
The term xe2x80x9cwherein two R1 moieties (or two Y moieties) may together form a 5 or 6 membered saturated or unsaturated ringxe2x80x9d is used herein to mean the formation of a napthylene ring system or a phenyl moiety having attached a 6 membered partially unsaturated ring such as a C6 cycloalkenyl, i.e hexene, or a C5 cyloalkenyl moiety, cyclopentene.
It is recognized that the compounds of the present invention may exist as stereoisomers, regioisomers, or diastereiomers. These compounds may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All of these compounds are included within the scope of the present invention.
The compounds of Formula (I) may be obtained by applying synthetic procedures, some of which are illustrated in the Schemes below. The synthesis provided for in these Schemes is applicable for the producing compounds of Formula (I) having a variety of different R, R1, and Aryl groups which are reacted, employing optional substituents which are suitably protected, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, in those cases, then affords compounds of the nature generally disclosed. Once the urea nucleus has been established, further compounds of these formulas may be prepared by applying standard techniques for functional group interconversion, well known in the art. While the schemes are shown with compounds only of Formula (I) this is merely for illustration purposes only.
Methods of Preparation
The title compounds can be synthesized from the thiouronium salt (2, Scheme 1). Rxe2x80x2 represents the (R13R14)v-HET linkage as defined in compounds of Formula (I). Z is also as defined in compounds of Formula (I). For purposes of illustration herein the schemes represent the W term by a substituted phenyl. 
The thiouronium salt(2, Scheme 1) can be synthesized by reacting sodium cyanamide with a commercially available isothiocyanate 1 (if the isothiocyanate is not commercially available it can be synthesized by reacting the desired amine with thiophosgene in the presence of a base like sodium bicarbonate). The thiouronium salt (2) can then be condensed with the appropriate substituted aniline in the presence of a coupling reagent like EDC.HCl or acidified to form the cyanothiourea and then reacted.
Alternatively 4 can be synthesized by reacting the commercially available (Aldrich Chem. Co.), diphenyl cyanocarboimidate (5, Scheme 2) with an amine to form an intermediate O-phenylisourea 6 which then reacts with the appropriately substituted aniline in the presence of trimethyl aluminum by the method of Atwal. (Atwal, K. S., Tetrahedron Lett, 35, 8085 (1994).). Compounds wherein Rxe2x80x2 is alkyl may be prepared by heating with the appropriately substituted alkylamine, but without the presence of catalyst. 
Alternatively the title compound can be synthesized using a protected ortho-substituted aniline (8, Scheme 3, See synthesis as described in U.S. provisional application U.S. Ser. No. 60/020655 filed Jun. 27, 1996, WO96/25157 filed Aug. 22, 1996, Widdowson et al.; and U.S. Ser. No. 08/701,299 filed Aug. 22, 1996 whose disclosures are incorporated herein by reference in their entirety. An ortho-substituted aniline (7, Scheme 3) is first protected (ie. tert-butyl dimethyl silyl, allyl, benzyl, mom or other suitable protecting group) by reacting the ortho-substituted aniline with the appropriate alkyl or silyl halide in the presence of a suitable base (ie. cesium carbonate, potassium carbonate or imidazole) in an aprotic solvent. The protected ortho-substituted aniline may also be synthesized from an ortho-substituted nitrobenzene (9) by reacting it with a protecting group under conditions well known in the art (see Greene, T Protecting Groups in Organic Synthesis, Wiley and Sons, New York, 1981) This protected ortho-substituted nitro compound is then reduced to the corresponding aniline using SnCl2 in EtOH or alternately H2/Pd or LiAlH4 in an aprotic solvent. 
This protected ortho-substituted aniline (8) can then be converted into an isothiocyanate using thiophosgene and then reacted with the anion ZNHxe2x88x92 (formed from reaction of ZNH2 with a base such as NaH). Z is as defined in compounds of Formula (I).The resulting thioanion can then be alkylated with an alkylating agent like methyl iodide to form a thioimidate such as 10 (Scheme 4). 
The thioimidate (10, Scheme 5) can be converted to the title compound 4 by reaction with the amine Rxe2x80x2NH2. This reaction can be accelerated by the addition of metal salt with a high affinity for sulfur such as mercuric oxide or silver acetate or by oxidation of the sulfur with dimethyloxirane to form a better leaving group. Finally the phenol protection is removed by standard methods to form the title compound 4. 
Alternately the title compound could be synthesized by reaction of a protected carbodiimide (11, Scheme 6) with the anion NHxe2x80x94Z (formed from reaction of NH2Z with a base such as NaH) or the neutral species NH2Z (Zxe2x95x90CN) and a tertiary amine base, such as Hxc3xcnig""s base (diisopropylethylamine), triethylamine, tri-isopropylethylamine, N,N-dimethylbenzylamine, or N,N-dimethylisopropylamine, under conditions where the nucleophile is present in large excess and the reaction time is kept as short as possible by carefully monitoring the reaction for completion followed by deprotection. Other suitable bases for use herein include, secondary amine, such as pyridine, and amino substituted pyridine derivatives. Suitable solvents for use herein when Z is cyano include various aprotic solvents, such as acetonitrile; halogenated solvents, such as chloroform and methylene chloride; ethyl gylcol-dimethyl ether (monoGLYME), dioxane, DMF and DMSO; or mixtures thereof, preferably acetonitrile. It is recognized by the skilled artisan that the limiting feature fo use of solvents herein will be the solubility of the cyano derivatived compound. For compounds wherein Z is other than cyano, while aprotic solvents are preferred, it is recognized by the skilled artisan that other suitable solvents, such as protic solvents, i.e. alcohols, may be used.
Preferably the reaction (when Z is cyano) temperature is from about xe2x88x9210xc2x0 to about 100xc2x0, preferably about 10xc2x0 to about 50xc2x0, more preferably around room temperature, i.e. 20 to 30xc2x0 C.
The protected Rxe2x80x3 moiety may be suitably deprotected using art recognized techniques. Preferably the deprotection is by deallylation catalyzed by palladium (O) when the protecting group is an allyl derivative. 
The carbodiimide 11 is prepared from the thiourea (12a, Scheme 7) by treatment with phosgene and a tertiary amine base or from the thiourea (12a) or urea (12b) by reaction with triphenylphosphine, carbon tetrachloride and triethylamnine.
The carbodiimide may also be prepared by reaction of the thiourea (12b) with an excess, such as 2 or more equivalents of methanesulfonyl chloride and a tertiary amine base, such as Hxc3xcnig""s base (diisopropylethylamine), triethylamine, tri-isopropylethylamine, N,N-dimethylbenzylamine, or N,N-dimethylisopropylamine, preferably triethylamine. The reaction may use any halogenated solvent, such as methylene chloride, chloroform, or tetrachloroethylene, etc.; suitable reaction temperatures are from about. xe2x88x9230xc2x0 C. to about 80xc2x0 C., preferably xe2x88x9210xc2x0 C. to about 50xc2x0 C., more preferably from about 0xc2x0 C. to about room temperature. See Fell and Coppola (Fell, J. B., Coppola, J. B., Syn Communications 25, 43, (1995). 
The thiourea or urea derivative may be synthesized as described in the U.S. provisional application U.S. Ser. No. 60/020655 and WO96/25157, supra. The thiourea (12a, Scheme 8) may also be prepared by reaction of the protected ortho-substituted aniline and two equivalents of an appropriate base such as NaH, KH, calcium hydride, and reacting this anion with a commercially available isothiocyanate (W1-NCS, wherein W1 is as defined for compounds of Formula (I)). The reaction make take place in any suitable aprotic solvent or halogenated solvent, preferably dimethyl formamide. Suitable reaction temperatures for this reaction are from about xe2x88x9210xc2x0 to about 50xc2x0.
If the desired isothiocyanate is not commercially available, it may be prepared by reaction of a corresponding aniline with thiophosgene and a suitable base such as sodium bicarbonate. 
Another aspect of the present invention are the novel compounds of Formula (II)
W1xe2x80x94Nxe2x95x90Cxe2x95x90Nxe2x80x94HETxe2x80x83xe2x80x83(II)
wherein W, and HET are as defined for formula (I), and W contains a protected or unprotected R group (Rxe2x80x3) as defined for compounds of Formula (I).
Another aspect of the present invention are the novel compounds of Formula (III)
W1xe2x80x94NHxe2x80x94C(S)xe2x80x94NHxe2x80x94HETxe2x80x83xe2x80x83(III)
wherein W, and HET are as defined for formula (I), and W contains a protected or unprotected R group (Rxe2x80x3) as defined for compounds of Formula (I).
Also, the protected (Rxe2x80x3) versions of compounds of Formula (I) are contemplated as being within the scope of this invention.
It is recognized that the guanidine functionality may hve a number of different tautomers, such as Wxe2x80x94Nxe2x80x94C(xe2x95x90NZ)xe2x80x94Nxe2x80x94HET; ZNxe2x95x90C(Nxe2x80x94HET)xe2x80x94Nxe2x80x94W; HETxe2x80x94Nxe2x80x94C(xe2x95x90NW)xe2x80x94NZ, all of which are within the scope of this invention.
Pharmaceutically acceptable salts of compounds of Formula (I) may be obtained in known manner, for example by treatment thereof with an appropriate amount of acid or base in the presence of a suitable solvent.
Method of Treatment
The compounds of Formula (I), or a pharmaceutically acceptable salt thereof can be used in the manufacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human, or other mammal, which is exacerbated or caused by excessive or unregulated IL-8 cytokine production by such mammal""s cell, such as but not limited to monocytes and/or macrophages, or other chemokines which bind to the IL-8 a or b receptor, also referred to as the type I or type II receptor.
Accordingly, the present invention provides a method of treating a chemokine mediated disease, wherein the chemokine is one which binds to an IL-8 a or b receptor and which method comprises administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In particular, the chemokines are IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2.
The compounds of Formula (I) are administered in an amount sufficient to inhibit cytokine function, in particular IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2, such that they are biologically regulated down to normal levels of physiological function, or in some case to subnormal levels, so as to ameliorate the disease state. Abnormal levels of IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2 for instance in the context of the present invention, constitute: (i) levels of free IL-8 greater than or equal to 1 picogram per mL; (ii) any cell associated IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2 above normal physiological levels; or (iii) the presence of IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2 above basal levels in cells or tissues in which IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2 respectively, is produced.
There are many disease states in which excessive or unregulated IL-8 production is implicated in exacerbating and/or causing the disease. Chemokine mediated diseases include psoriasis, atopic dermatitis, arthritis, asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, inflammatory bowel disease, Crohn""s disease, ulcerative colitis, stroke, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, cardiac and renal reperfusion injury, glomerulonephritis, thrombosis, graft vs. host reaction, alzheimers disease, allograft rejections, malaria, restinosis, angiogenesis or undesired hematopoietic stem cells release.
These diseases are primarily characterized by massive neutrophil infiltration, T-cell infiltration, or neovascular growth, and are associated with increased IL-8, GROxcex1, GROxcex2, GROxcex3or NAP-2 production which is responsible for the chemotaxis of neutrophils into the inflammatory site or the directional growth of endothelial cells. In contrast to other inflammatory cytokines (IL-1, TNF, and IL-6), IL-8, GROxcex1, GROxcex2, GROxcex3 or NAP-2 has the unique property of promoting neutrophil chemotaxis, enzyme release including but not limited to elastase release as well as superoxide production and activation. The xcex1-chemokines but particularly, GROxcex1, GROxcex2, GROxcex3 or NAP-2, working through the IL-8 type I or II receptor can promote the neovascularization of tumors by promoting the directional growth of endothelial cells. Therefore, the inhibition of IL-8 induced chemotaxis or activation would lead to a direct reduction in the neutrophil infiltration.
Recent evidence also implicates the role of chemokines in the treatment of HIV infections, Littleman et al., Nature 381, pp661 (1996) and Koup et al., Nature 381, pp 667 (1996).
The present invention also provides for a means of treating, in an acute setting, as well as preventing, in those individuals deemed susceptible to, CNS injuries by the chemokine receptor antagonist compounds of Formula (I).
CNS injuries as defined herein include both open or penetrating head trauma, such as by surgery, or a closed head trauma injury, such as by an injury to the head region. Also included within this definition is ischemic stroke, particularly to the brain area.
Ischemic stroke may be defined as a focal neurologic disorder that results from insufficient blood supply to a particular brain area, usually as a consequence of an embolus, thrombi, or local atheromatous closure of the blood vessel. The role of inflammatory cytokines in this are has been emerging and the present invention provides a mean for the potential treatment of these injuries. Relatively little treatment, for an acute injury such as these has been available.
TNF-xcex1 is a cytokine with proinflammatory actions, including endothelial leukocyte adhesion molecule expression. Leukocytes infiltrate into ischemic brain lesions and hence compounds which inhibit or decrease levels of TNF would be useful for treatment of ischemic brain injury. See Liu et al., Stoke, Vol. 25., No. 7, pp 1481-88 (1994) whose disclosure is incorporated herein by reference.
Models of closed head injuries and treatment with mixed 5-LO/CO agents is discussed in Shohami et al., J. of Vaisc and Clinical Physiology and Pharmacology, Vol. 3, No. 2, pp. 99-107 (1992) whose disclosure is incorporated herein by reference. Treatment which reduced edema formation was found to improve functional outcome in those animals treated.
The compounds of Formula (I) are administered in an amount sufficient to inhibit IL-8, binding to the IL-8 alpha or beta receptors, from binding to these receptors, such as evidenced by a reduction in neutrophil chemotaxis and activation. The discovery that the compounds of Formula (I) are inhibitors of IL-8 binding is based upon the effects of the compounds of Formulas (I) in the in vitro receptor binding assays which are described herein. The compounds of Formula (I) have been shown, in some instances, to be dual inhibitors of both recombinant type I and type II IL-8 receptors. Preferably the compounds are inhibitors of only one receptor, more preferably Type II.
As used herein, the term xe2x80x9cIL-8 mediated disease or disease statexe2x80x9d refers to any and all disease states in which IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2 plays a role, either by production of IL-8, GROxcex1, GROxcex2, GROxcex3 or NAP-2 themselves, or by IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2 causing another monokine to be released, such as but not limited to IL-1, IL-6 or TNF. A disease state in which, for instance, IL-1 is a major component, and whose production or action, is exacerbated or secreted in response to IL-8, would therefore be considered a disease stated mediated by IL-8.
As used herein, the term xe2x80x9cchemokine mediated disease or disease statexe2x80x9d refers to any and all disease states in which a chemokine which binds to an IL-8 a or b receptor plays a role, such as but not limited to IL-8, GROxcex1, GROxcex2, GROxcex3, ENA-78 or NAP-2. This would include a disease state in which, IL-8 plays a role, either by production of IL-8 itself, or by IL-8 causing another monokine to be released, such as but not limited to IL-1, IL-6 or TNF. A disease state in which, for instance, IL-1 is a major component, and whose production or action, is exacerbated or secreted in response to IL-8, would therefore be considered a disease stated mediated by IL-8.
As used herein, the term xe2x80x9ccytokinexe2x80x9d refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response. A cytokine includes, but is not limited to, inonokines and lymphokines, regardless of which cells produce them. For instance, a monokine is generally referred to as being produced and secreted by a mononuclear cell, such as a macrophage and/or monocyte. Many other cells however also produce monokines, such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epideral keratinocytes and B-lymphocytes. Lymphokines are generally referred to as being produced by lymphocyte cells. Examples of cytokines include, but are not limited to, Interleukin-1 (IL-1), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Tumor Necrosis Factor-alpha (TNF-xcex1) and Tumor Necrosis Factor beta (TNF-xcex2).
As used herein, the term xe2x80x9cchemokinexe2x80x9d refers to any secreted polypeptide that affects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response, similar to the term xe2x80x9ccytokinexe2x80x9d above. A chemokine is primarily secreted through cell transmembranes and causes chemotaxis and activation of specific white blood cells and leukocytes, neutrophils, monocytes, macrophages, T-cells, B-cells, endothelial cells and smooth muscle cells. Examples of chemokines include, but are not limited to, IL-8, GRO-xcex1, GRO-xcex2, GROxcex3, ENA-78, NAP-2, IP-10, MIP-1xcex1, MIP-xcex2, PF4, and MCP 1, 2, and 3.
In order to use a compound of Formula (I) or a pharmaceutically acceptable salt thereof in therapy, it will normally be formulated into a pharmaceutical composition in accordance with standard pharmaceutical practice. This invention, therefore, also relates to a pharmaceutical composition comprising an effective, non-toxic amount of a compound of Formula (I) and a pharmaceutically acceptable carrier or diluent.
Compounds of Formula (I), pharmaceutically acceptable salts thereof and pharmaceutical compositions incorporating such may conveniently be administered by any of the routes conventionally used for drug administration, for instance, orally, topically, parenterally or by inhalation. The compounds of Formula (I) may be administered in conventional dosage forms prepared by combining a compound of Formula (I) with standard pharmaceutical carriers according to conventional procedures. The compounds of Formula (I) may also be administered in conventional dosages in combination with a known, second therapeutically active compound. These procedures may involve mixing, granulating and compressing or dissolving the ingredients as appropriate to the desired preparation. It will be appreciated that the form and character of the pharmaceutically acceptable character or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. The carrier(s) must be xe2x80x9cacceptablexe2x80x9d in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
The pharmaceutical carrier employed may be, for example, either a solid or liquid. Exemplary of solid carriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary of liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the carrier or diluent may include time delay material well known to the art, such as glyceryl mono-stearate or glyceryl distearate alone or with a wax.
A wide variety of pharmaceutical forms can be employed. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge. The amount of solid carrier will vary widely but preferably will be from about 25 mg. to about 1 g. When a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or nonaqueous liquid suspension.
Compounds of Formula (I) may be administered topically, that is by non-systemic administration. This includes the application of a compound of Formula (I) externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.
Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. The active ingredient may comprise, for topical administration, from 0.001% to 10% w/w, for instance from 1% to 2% by weight of the Formulation. It may however comprise as much as 10% w/w but preferably will comprise less than 5% w/w, more preferably from 0.1% to 1% w/w of the Formulation.
Lotions according to the present invention include those suitable for application to the skin or eye. An eye lotion may comprise a sterile aqueous solution optionally containing a bactericide and may be prepared by methods similar to those for the preparation of drops. Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and/or a moisturizer such as glycerol or an oil such as castor oil or arachis oil.
Creams, ointments or pastes according to the present invention are semi-solid formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution or suspension in an aqueous or non-aqueous fluid, with the aid of suitable machinery, with a greasy or non-greasy base. The base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives or a fatty acid such as steric or oleic acid together with an alcohol such as propylene glycol or a macrogel. The formulation may incorporate any suitable surface active agent such as an anionic, cationic or non-ionic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof. Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such as lanolin, may also be included.
Drops according to the present invention may comprise sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal and/or fungicidal agent and/or any other suitable preservative, and preferably including a surface active agent. The resulting solution may then be clarified by filtration, transferred to a suitable container which is then sealed and sterilized by autoclaving or maintaining at 98-100xc2x0 C. for half an hour. Alternatively, the solution may be sterilized by filtration and transferred to the container by an aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%). Suitable solvents for the preparation of an oily solution include glycerol, diluted alcohol and propylene glycol.
Compounds of formula (I) may be administered parenterally, that is by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. Appropriate dosage forms for such administration may be prepared by conventional techniques. Compounds of Formula (I) may also be administered by inhalation, that is by intranasal and oral inhalation administration. Appropriate dosage forms for such administration, such as an aerosol formulation or a metered dose inhaler, may be prepared by conventional techniques.
For all methods of use disclosed herein for the compounds of Formula (I), the daily oral dosage regimen will preferably be from about 0.01 to about 80 mg/kg of total body weight. The daily parenteral dosage regimen about 0.001 to about 80 mg/kg of total body weight. The daily topical dosage regimen will preferably be from 0.1 mg to 150 mg, administered one to four, preferably two or three times daily. The daily inhalation dosage regimen will preferably be from about 0.01 mg/kg to about 1 mg/kg per day. It will also be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formula (I) or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, i.e., the number of doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests.